一种改良型的免疫印迹法  

A New Method of Western Blot to Economize Antibodies

作　　者：尚蕾[1] 黄铠[2] 曹妍群[1,2] 黄菊芳[1] 

机构地区：[1]中南大学湘雅医学院人体解剖与神经生物学系,湖南长沙410011 [2]邵阳医学高等专科学校,湖南邵阳422000

出　　处：《现代生物医学进展》2012年第32期6368-6370,共3页Progress in Modern Biomedicine

基　　金：国家自然科学基金(81070729);中南大学研究生学位论文创新基金(2010ssxt258);高等学校博士学科点专项科研基金(20100162110067)

摘　　要：目的:蛋白免疫印迹法是现代生物实验过程中运用最为广泛的实验技术,常规的免疫印迹法在应用过程中存在很多弊端,如浪费抗体等,因此非常有必要探索出一种新型的免疫印迹法,本文旨在探索一种能够节约抗体的免疫印迹实验方法。方法:将8只SD大鼠随机分常规组和改进组两组,每组4只,活取视网膜组织,进行组织匀浆、蛋白定量,取不同蛋白总量的匀浆变性液进行β-Tubulin的免疫印迹实验,比较两组之间β-Tubulin的蛋白表达量之间是否存在显著性差异,实验需重复三次。结果:不同总量蛋白的免疫印迹显示两组之间β-Tubulin的表达量并无显著性差异。但是,相比常规方法,改进法使用的抗体量更少,条带更容易检测得到。结论:改进后的免疫印迹法能有效的节约抗体,操作方便,实用性强。Objective： Westem blotting is an extensive applied technology in our morden biology research work. There are a plenty of malpractice in conventional methods, such as waste antibodies, waste a lot of time. Therefore it is so important to explore a new method of western blot to conquer the defects in common method. Our purpose is to explore a new method of western blot to save anti-bodies. Methods： The eight Sprague-Dawley rats, were randomly divided into two groups named conventional group and improved gourp. Eyeballs were removed immediately from deeply anesthetized rats on ice, Retinas were homogenized by digest solution and pro-tein concentration were determined by Bicinnchoninic acid, then we continued the westem blot process in two different protein concentrations using β-Tubulin antibody. Every expriment repeats three times. Results： Comparing conventional groups to improved groups, no significant differences were found in the protein expression of β-Tubulin. However, less antibodies were used and strips were more clear on films in improved groups. Conclusion： Our new method may help researches to save a plenty of antibodies, the method may became more effectively, and could be easy handled.

关 键 词：WESTERN BLOT Β-TUBULIN 视网膜 抗体 

分 类 号：R392[医药卫生—免疫学]